title: Single cell and spheroid collagen type I invasion assay.
abstract: Tumor invasion is the outcome of a complex interplay between cancer cells and the stromal environment and requires the infiltration of a dense, cross-linked meshwork of collagen type I extracellular matrix. We use a membrane-free single-cell and spheroid-based complementary model to study cancer invasion through native collagen type I matrices. Cell morphology is preserved during the assays allowing real-time monitoring of invasion-induced changes in cell structure and F-actin organization. Combination of these models with computerized quantification permits the calculation of highly reproducible and operator-independent data. These assays are versatile in the use of fluorescent probes and have a flexible kinetic endpoint. Once the optimal experimental conditions are empirically determined, the collagen type I invasion assays can be used for preclinical validation of small-molecule inhibitors targeting invasion. Initiation and monitoring of the single-cell and spheroid invasion model can be achieved in 8 h (over 3 days) and in 14 h (over 5 days), respectively.
authors.affiliation: Department of Radiotherapy and Nuclear Medicine, Laboratory of Experimental Cancer Research, Ghent University Hospital, Ghent, Belgium.
authors.forename: Olivier
authors.initials: O
authors.lastname: De Wever
authors.forename: An
authors.initials: A
authors.lastname: Hendrix
authors.forename: Astrid
authors.initials: A
authors.lastname: De Boeck
authors.forename: Frank
authors.initials: F
authors.lastname: Eertmans
authors.forename: Wendy
authors.initials: W
authors.lastname: Westbroek
authors.forename: Geert
authors.initials: G
authors.lastname: Braems
authors.forename: Marc E
authors.initials: ME
authors.lastname: Bracke
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D024042:Collagen Type I / Q000494:pharmacology*; D006801:Humans; D008954:Models, Biological; D051381:Rats; D059010:Single-Cell Analysis / Q000379:methods*; D018874:Spheroids, Cellular / Q000166:cytology*
chemical_list: D024042:Collagen Type I